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Plasmid Files

pEX-C-GST

PrecisionShuttle™ bacterial vector for inducible expression of a protein with a cleavable C-terminal GST tag.

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pEX-C-GST.dna
Map and Sequence File:    Download    Open   
Sequence Author:  OriGene
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AanI - PsiI (5181) DraIII (5056) BsaAI (5053) BsaI (3965) AhdI (3904) AlwNI (3427) BamHI (2882) PluTI (2532) EcoRI (184) BsaBI * - BseRI - BspDI * - ClaI * (288) NheI (329) BmtI (333) RsrII (335) MluI (341) BsiWI (344) NotI (350) AvaI - BsoBI - PaeR7I - PspXI - XhoI (356) BmeT110I (357) EcoNI (393) MscI (590) BstBI (780) SwaI (810) AsiSI - SgfI (1083) MreI - SgrAI (1085) PmeI (1118) FseI (1128) BlpI (1158) StyI (1180) BtgI (1236) SacII (1239) PflMI (1471) BstAPI (1572) BstEII (2070) PspOMI (2096) ApaI (2100) HincII - HpaI (2395) BsmBI (2504) KasI (2528) NarI * (2529) SfoI (2530) pEX-C-GST 5253 bp
AanI  (5181)
1 site
T T A T A A A A T A T T
PsiI  (5181)
1 site
T T A T A A A A T A T T
DraIII  (5056)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
BsaAI  (5053)
1 site
Y A C G T R R T G C A Y
BsaI  (3965)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
AhdI  (3904)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
AlwNI  (3427)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
BamHI  (2882)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
PluTI  (2532)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
EcoRI  (184)
1 site
G A A T T C C T T A A G
BsaBI  (288)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BseRI  (288)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
BspDI  (288)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (288)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
NheI  (329)
1 site
G C T A G C C G A T C G
BmtI  (333)
1 site
G C T A G C C G A T C G
RsrII  (335)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
MluI  (341)
1 site
A C G C G T T G C G C A
BsiWI  (344)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
NotI  (350)
1 site
G C G G C C G C C G C C G G C G
AvaI  (356)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (356)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
PaeR7I  (356)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (356)
1 site
V C T C G A G B B G A G C T C V
XhoI  (356)
1 site
C T C G A G G A G C T C
BmeT110I  (357)
1 site
C Y C G R G G R G C Y C
EcoNI  (393)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
MscI  (590)
1 site
T G G C C A A C C G G T
BstBI  (780)
1 site
T T C G A A A A G C T T
SwaI  (810)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
AsiSI  (1083)
1 site
G C G A T C G C C G C T A G C G
SgfI  (1083)
1 site
G C G A T C G C C G C T A G C G
MreI  (1085)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (1085)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
PmeI  (1118)
1 site
G T T T A A A C C A A A T T T G
FseI  (1128)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
BlpI  (1158)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
StyI  (1180)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BtgI  (1236)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
SacII  (1239)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
PflMI  (1471)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
BstAPI  (1572)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
BstEII  (2070)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
PspOMI  (2096)
1 site
G G G C C C C C C G G G
ApaI  (2100)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
HincII  (2395)
1 site
G T Y R A C C A R Y T G
HpaI  (2395)
1 site
G T T A A C C A A T T G
BsmBI  (2504)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
KasI  (2528)
1 site
G G C G C C C C G C G G
NarI  (2529)
1 site
G G C G C C C C G C G G
* Blocked by Dcm methylation.
Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (2530)
1 site
G G C G C C C C G C G G
lacI
1539 .. 2621  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lacI
1539 .. 2621  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
AmpR
3831 .. 4691  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3831 .. 4622  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3831 .. 4691  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4623 .. 4691  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3831 .. 4691  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
GST
383 .. 1036  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
GST
383 .. 1036  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
ori
3072 .. 3660  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3072 .. 3660  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
4823 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
4823 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
AmpR promoter
4692 .. 4796  =  105 bp
AmpR promoter
4692 .. 4796  =  105 bp
lacI promoter
1461 .. 1538  =  78 bp
lacI promoter
1461 .. 1538  =  78 bp
MCS
286 .. 361  =  76 bp
multiple cloning site
MCS
286 .. 361  =  76 bp
multiple cloning site
T7 terminator
1169 .. 1215  =  47 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
1169 .. 1215  =  47 bp
transcription terminator for bacteriophage T7 RNA polymerase
lac operator
235 .. 259  =  25 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
235 .. 259  =  25 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
TEV Site
362 .. 382  =  21 bp
7 amino acids  =  870.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV Site
362 .. 382  =  21 bp
7 amino acids  =  870.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV Site
1058 .. 1078  =  21 bp
7 amino acids  =  870.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV Site
1058 .. 1078  =  21 bp
7 amino acids  =  870.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
T7 promoter
216 .. 234  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
216 .. 234  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
279 .. 284  =  6 bp
ribosome binding site
RBS
279 .. 284  =  6 bp
ribosome binding site
ORF:  3961 .. 4227  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  383 .. 1114  =  732 bp
ORF:  243 amino acids  =  28.1 kDa
ORF:  1472 .. 1789  =  318 bp
ORF:  105 amino acids  =  11.2 kDa
ORF:  2378 .. 2641  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  2744 .. 2971  =  228 bp
ORF:  75 amino acids  =  8.5 kDa
ORF:  1662 .. 2621  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  820 .. 1047  =  228 bp
ORF:  75 amino acids  =  8.3 kDa
ORF:  2404 .. 2655  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  3831 .. 4691  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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